Development and implementation of a tool for measuring the training effectiveness of the patient-centered consultation model.
Background: The patient-centered consultation model comprises four elements: exploring health, illness and disease experiences, understanding the whole person, finding common ground, and enhancing the patient-doctor relationship. This method is taught at the course in general practice at Copenhagen University. The aim of the study was to develop a simple tool consisting of a questionnaire about the patient-centered elements and a test video consultation. The outcome is the change in the students' ability to identify these elements. Used as a pre-course and post-course test it can inform the teachers which elements of the patient-centered consultation need intensifying in the teaching. Methods: The students from a course in general practice volunteered to participate in all steps of the development. They took part in individual interviews to select items from an already existing questionnaire (DanSCORE). The preliminary questionnaire was tested for face and content validity, pilot-tested and tested for test-retest reliability. All video consultations were transcribed and assessed for patient-centered elements through a conversation analysis. The videos showed medical students seeing real patients. Results: The preliminary version of the questionnaire (called DanOBS) had 23 items. In the subsequent interviews, items were reduced to 17, each with three response options. After a pilot test, the questionnaire was further reduced to 13 items, all strictly relevant to the model and with two response options. The final questionnaire had acceptable test-retest reliability. The number of test consultation videos underwent a reduction from six videos to one. Conclusions: The DanOBS combined with a test video consultation, used as a pre-and post-course test demonstrates for teachers which elements in the patient-centered consultation need to be intensified in the teaching.


Sequence analysis of the S1 glycoprotein of infectious bronchitis viruses: identification of a novel genotypic group in Australia.
Sequencing of the S1 genes of nine Australian strains of infectious bronchitis virus (IBV) identified two genotypically distinct groups of strains. The strains Vic S, V5/90, N1/62, N3/62, N9/74, and N2/75 comprised group I, sharing 80.7-98.3% identity in their deduced amino acid sequences. All group I strains were able to replicate in the trachea and kidney but only four strains, Vic S, N1/62, N9/74, and N2/75, were nephropathogenic, the latter three causing mortalities ranging from 32 to 96%. Group II contained strains N1/88, Q3/88 and V18/91 which only replicated in the trachea, inducing no mortalities. These viruses showed 72.3-92.8% amino acid identity to each other and only 53.8-61.7% identity to viruses of the first group. They were also distinct from the Massachusetts 41 and D1466 strains (47.5-55.7% amino acid identity). Thus N1/88, Q3/88 and V18/91 form a new group of viruses which are genotypically distinct from all previously characterized IBV strains. No definite correlations were established between the S1 amino acid sequences and the nephropathogenicity of strains.


Posttranslational modification of CENP-A influences the conformation of centromeric chromatin.
Centromeres are chromosomal loci required for accurate segregation of sister chromatids during mitosis. The location of the centromere on the chromosome is not dependent on DNA sequence, but rather it is epigenetically specified by the histone H3 variant centromere protein A (CENP-A). The N-terminal tail of CENP-A is highly divergent from other H3 variants. Canonical histone N termini are hotspots of conserved posttranslational modification; however, no broadly conserved modifications of the vertebrate CENP-A tail have been previously observed. Here, we report three posttranslational modifications on human CENP-A N termini using high-resolution MS: trimethylation of Gly1 and phosphorylation of Ser16 and Ser18. Our results demonstrate that CENP-A is subjected to constitutive initiating methionine removal, similar to other H3 variants. The nascent N-terminal residue Gly1 becomes trimethylated on the α-amino group. We demonstrate that the N-terminal RCC1 methyltransferase is capable of modifying the CENP-A N terminus. Methylation occurs in the prenucleosomal form and marks the majority of CENP-A nucleosomes. Serine 16 and 18 become phosphorylated in prenucleosomal CENP-A and are phosphorylated on asynchronous and mitotic nucleosomal CENP-A and are important for chromosome segregation during mitosis. The double phosphorylation motif forms a salt-bridged secondary structure and causes CENP-A N-terminal tails to form intramolecular associations. Analytical ultracentrifugation of phospho-mimetic CENP-A nucleosome arrays demonstrates that phosphorylation results in greater intranucleosome associations and counteracts the hyperoligomerized state exhibited by unmodified CENP-A nucleosome arrays. Our studies have revealed that the major modifications on the N-terminal tail of CENP-A alter the physical properties of the chromatin fiber at the centromere.


Pericardial Tamponade Caused by Rotational Atherectomy Ablation Grinding Through a Barrier Stent Protruding From the Diagonal Artery: A Case Report.
Rotational atherectomy (RA) has been described in case reports to be effective in ablating under-expanded stents. We present a case of calcified bifurcation lesion percutaneous coronary intervention (PCI), in which the branch stent protruding into main branch blocked the balloon getting through, and the following RA led to pericardial tamponade. The first stent was deployed from the proximal of left anterior descending artery (LAD) to the proximal of diagonal for an 80-year-old man who suffered from non-ST-elevation myocardial infarction to make a Culotte dual stent PCI. The operator failed to make balloon get through the struts protruding into LAD lumen anyway. After RA drill through the LAD lumen, it showed an excellent angiographic outcome. Stent deployment in LAD was successful, but pericardial tamponade occurred in a short time. After pericardiocentesis was performed under fluoroscopic guidance, the patient was uneventful. After 1 year, the follow-up coronary computed tomography (CT) angiography showed no in-stent restenosis, stent fracture or disrupted struts protruding outside of the vessel's outline of LAD and the first diagonal (D1). This case shows stents' RA could be dangerous in grinding across the stents' lateral hole in an unsymmetrical lumen. Although RA could be a useful remedy in the situation of under-expansion of implanted stents, the debulking should be performed for longitudinal stent ablation and cautiously performed for bifurcation lesion in which the protruding stents from side branch were unsymmetrically blocking the path.


Axons: isolation from mammalian central nervous system.
Centrifugation of a homogenate of white matter, in a solution of buffered sucrose containing salt, produces a floating layer of myelinated axons. When these are suspended in hypotonic buffer, the mnyelin swells and strips away from the axon. Axons are then separated from the myelin by centrifugation. The resulting preparation consists of a variable population of processes with lengths up to 200 micrometers and diameters between 0.3 and 5.0 micrometers. The axons contain neurofilaments and mitochondria, although no axolemma or neurotubules are evident. The preparation contains cerebroside and sulfatide, yet is essentially free of myelin.


Acute systemic DNA damage in youth does not impair immune defense with aging.
Aging-related decline in immunity is believed to be the main driver behind decreased vaccine efficacy and reduced resistance to infections in older adults. Unrepaired DNA damage is known to precipitate cellular senescence, which was hypothesized to be the underlying cause of certain age-related phenotypes. Consistent with this, some hallmarks of immune aging were more prevalent in individuals exposed to whole-body irradiation (WBI), which leaves no anatomical repository of undamaged hematopoietic cells. To decisively test whether and to what extent WBI in youth will leave a mark on the immune system as it ages, we exposed young male C57BL/6 mice to sublethal WBI (0.5-4 Gy), mimicking human survivor exposure during nuclear catastrophe. We followed lymphocyte homeostasis thorough the lifespan, response to vaccination, and ability to resist lethal viral challenge in the old age. None of the irradiated groups showed significant differences compared with mock-irradiated (0 Gy) animals for the parameters measured. Even the mice that received the highest dose of sublethal WBI in youth (4 Gy) exhibited equilibrated lymphocyte homeostasis, robust T- and B-cell responses to live attenuated West Nile virus (WNV) vaccine and full survival following vaccination upon lethal WNV challenge. Therefore, a single dose of nonlethal WBI in youth, resulting in widespread DNA damage and repopulation stress in hematopoietic cells, leaves no significant trace of increased immune aging in a lethal vaccine challenge model.


Syncope and ventricular arrhythmias in hypertrophic cardiomyopathy are not related to the derangement of coronary microvascular function.
Non-sustained ventricular tachycardia on Holter and syncope have been considered risk factors for sudden death in hypertrophic cardiomyopathy. In these patients the coronary vasodilator reserve is impaired despite normal coronaries, so we evaluated the correlation between the severity of coronary vasodilator reserve impairment and the occurrence of syncope and non-sustained ventricular tachycardia. Eighty-four patients with hypertrophic cardiomyopathy (62 males, age 43 +/- 12 years) had a two-dimensional echocardiographic study and a 48-h Holter. Myocardial blood flow was measured by positron emission tomography, at baseline and after dipyridamole, and the coronary vasodilator reserve was computed as dipyridamole myocardial blood flow/baseline myocardial blood flow. In 27 patients, subendocardial and subepicardial myocardial blood flow was measured in the septum and the subendocardial/subepicardial ratio was computed. Twenty of 84 patients had at least one syncopal episode, and 26 had at least one run of non-sustained ventricular tachycardia on Holter. Baseline and dipyridamole myocardial blood flow, coronary vasodilator reserve, and baseline and dipyridamole subendocardial/subepicardial myocardial blood flow ratio were similar in patients with and without syncope and with and without non-sustained ventricular tachycardia on Holter. However, patients with non-sustained ventricular tachycardia had larger left ventricular end-diastolic (47 +/- 6 vs 44 +/- 5 mm, P < 0.05) and end-systolic diameters (30 +/- 6 vs 27 +/- 4 mm, P < 0.05). (1) Coronary vasodilation is not more severely impaired in patients with hypertrophic cardiomyopathy and syncope or non-sustained ventricular tachycardia. (2) The left ventricle is more dilated in hypertrophic cardiomyopathy with non-sustained ventricular tachycardia.


Applying Inverse Probability Weighting to Measure Contraceptive Prevalence Using Data from a Community-Based Reproductive Health Intervention in Pakistan.
Many community-based reproductive health programs use their program data to monitor progress toward goals. However, using such data to assess programmatic impact on outcomes such as contraceptive use poses methodological challenges. Inverse probability weighting (IPW) may help overcome these issues. Data on 33,162 women collected in 2013-2015 as part of a large-scale community-based reproductive health initiative were used to produce population-level estimates of the contraceptive prevalence rate (CPR) and modern contraceptive prevalence rate (mCPR) among married women aged 15-49 in Pakistan's Korangi District. To account for the nonrandom inclusion of women in the sample, estimates of contraceptive prevalence during the study's four seven-month intervention periods were made using IPW; these estimates were compared with estimates made using complete case analysis (CCA) and the last observation carried forward (LOCF) method-two approaches for which modeling assumptions are less flexible. In accordance with intervention protocols, the likelihood that women were visited by intervention personnel and thus included in the sample differed according to their past and current contraceptive use. Estimates made using IPW suggest that the CPR increased from 51% to 64%, and the mCPR increased from 34% to 53%, during the study. For both outcomes, IPW estimates were higher than CCA estimates, were generally similar to LOCF estimates and yielded the widest confidence intervals. IPW offers a powerful methodology for overcoming estimation challenges when using program data that are not representative of the population in settings where cost impedes collection of outcome data for an appropriate control group.


Effect of different luting cements on fracture resistance in endodontically treated teeth.
The aim of the present study was to evaluate the effect of three types of luting cements used for post cementation on the fracture resistance of endodontically treated maxillary premolars, restored with resin composite. One hundred intact single-rooted human maxillary premolars were randomly divided into 5 groups of 20 each. In groups 2-5, post spaces were prepared after root canal treatment and clinical crown reduction up to 1.5 mm above the CEJ. Teeth were divided in groups as follows: Group 1: intact teeth, Group 2: active prefabricated metallic posts (PMP), Group 3: PMP cemented with zinc phosphate luting cement, Group 4: PMP cemented with glass ionomer luting cement and Group 5: PMP cemented with resin luting cement. In groups 2-5 the teeth were restored with resin composite. Following thermocycling, the palatal cusp of each specimen was loaded to compression at an angle of 150˚ to its longitudinal axis at a strain rate of 2 mm/min until fracture occurred. Data were analyzed using one-way ANOVA and a post hoc Tukey test. Chi-square test was used for comparison of failure mode. There were significant differences in fracture resistance between the test groups (P<0.001). The differences between group 2 with groups 1, 4 and 5 were statistically significant (P<0.05); whereas there were no significant differences in fracture resistance between the two other groups (P>0.05). Furthermore, there were no significant differences in the mode of failure between the 5 groups (P>0.05). Zinc phosphate, glass ionomer and resin luting cements showed similar behaviors and achieved fracture resistance comparable to intact teeth. However, the use of active post (without cement) adversely affected the fracture resistance of root canal treated teeth.


Are school vending machines loaded with calories and fat: an assessment of 106 middle and high schools.
The purpose of this study was to describe the extent to which vending offerings in 106 schools in the St. Paul-Minneapolis, Minnesota metropolitan area, met criteria for types of beverages, fat, and calories based on selected criteria offered by the Institute of Medicine. Schools where youth participants were attending for the 2006-2007 school year were identified and invited to participate in the study (n = 143); 81% of schools (n = 116) agreed to participate. Of the 116 schools, 106 had vending machines. Across schools with vending machines (n = 106), 5085 food and 8442 beverage items were offered. Overall, only 18% of beverage items met criteria for calories and type of beverage; significantly more items in public schools met the criteria as compared to private schools (19% vs 12%; p < .01). This difference was also significant for high schools as compared to middle schools (18% vs 22%; p < .01). For food items, 41% met calorie criteria, 45% met fat criteria, and 22% met both fat and calorie criteria. Significantly more food items met both criteria in public than private schools (22% vs 18%; p = .01), while high schools (22%) and middle schools (21%) were similar. A very small proportion of foods (< 5%) would have met the full criteria suggested by the Institute of Medicine for competitive foods. Overall, foods and beverages offered in vending machines continue to be high in fat and calories. Public schools are doing a slightly better job of providing healthy foods as compared to private schools.


The natural history of mountain cedar pollinosis.
During the winter months, pollen from the mountain cedar (MC) (Juniperus sabinoides) causes severe respiratory tract allergy in central Texas. We have been impressed with the fact that many of our MC-allergic patients had only allergic rhinitis and were only sensitive to MC pollen. We therefore studied 234 unselected MC-allergic patients at the end of the MC season. The main criteria for inclusion into the study was a bona fide history of MC pollinosis confirmed by a positive skin test. All patients completed a detailed history questionnaire, were prick skin tested by using 1:20 w/v, extracts, and had a total IgE determined with commercially available reagents (PRIST). Thirty-four percent of patients were found to be allergic only to MC and 66% were allergic to MC and other aeroallergens. As a group, patients allergic only to MC had significantly lower total IgE levels (84 IU/ml vs. 360, p < 0.001) (normal < 180), required much longer exposure intervals to cedar pollen before developing MC pollinosis (14.4 yr vs, 5.69, p < 0.001), had less of a family history of allergic disease (43% vs. 88%, p < 0.001), had less of a history of asthma or eczema (11% vs. 39%, p < 0.001), and developed allergic disease at a later age (39 yr vs. 13, p < 0.001) when compared to patients with multiple allergies. These results suggest that the MC pollen may be unique in causing allergic rhinitis in patients who have no other sensitivities. A possible explanation may lie in the carbohydrate nature of the main allergen of the MC pollen, which may facilitate allergen transport through the respiratory tract mucosa and subsequent sensitization.


Probe-sample optical interaction: size and wavelength dependence in localized plasmon near-field imaging.
The probe-sample optical interaction in apertureless near-field optical microscopy is studied at 633 nm and 808 nm excitation wavelengths using gold nanodisks as model systems. The near-field distributions of the dipolar and quadrupolar surface plasmon modes have been mapped successfully using metal coated probes with different polarization combinations of excitation and detection except when the incident and the scattered light polarizations are chosen to be parallel to the probe axis. For the parallel polarization of the incident and the scattered light, the pattern of the near-field distribution differs from the inherent plasmon mode structures of the sample, depending sensitively on the sample size and excitation energy. For a given excitation energy, the near-field amplitude shifts from one pole to the other as the sample size increases, having nearly equal amplitude at the two poles when the plasmon resonance peak spectrally overlaps with the excitation energy.


Love Conquers all but Nicotine: Spousal Peer Effects on the Decision to Quit Smoking.
If two partners smoke, their quit behavior may be related through correlation in unobserved individual characteristics and through common shocks. However, there may also be a causal effect whereby the quit behavior of one partner is affected by the quit decision of the other partner. If so, there is a spousal peer effect on the decision to quit smoking. We use data containing retrospective information of Dutch partnered individuals about their age of onset of smoking and their age of quitting smoking. We estimate mixed proportional hazard models of starting rates and quit rates of smoking in which we allow unobserved heterogeneity to be correlated across partners. Using a timing of events approach, we determine whether the quitting-to-smoke decision of one partner has a causal effect on the quitting-to-smoke decision of the other partner. We find no evidence of substantial spousal peer effects in the decision to quit smoking. Apparently, love conquers all but nicotine addiction. Copyright © 2017 John Wiley & Sons, Ltd.


Lenalidomide, Bortezomib, and Dexamethasone with Transplantation for Myeloma.
High-dose chemotherapy plus autologous stem-cell transplantation has been the standard treatment for newly diagnosed multiple myeloma in adults up to 65 years of age. However, promising data on the use of combination therapy with lenalidomide, bortezomib, and dexamethasone (RVD) in this population have raised questions about the role and timing of transplantation. We randomly assigned 700 patients with multiple myeloma to receive induction therapy with three cycles of RVD and then consolidation therapy with either five additional cycles of RVD (350 patients) or high-dose melphalan plus stem-cell transplantation followed by two additional cycles of RVD (350 patients). Patients in both groups received maintenance therapy with lenalidomide for 1 year. The primary end point was progression-free survival. Median progression-free survival was significantly longer in the group that underwent transplantation than in the group that received RVD alone (50 months vs. 36 months; adjusted hazard ratio for disease progression or death, 0.65; P<0.001). This benefit was observed across all patient subgroups, including those stratified according to International Staging System stage and cytogenetic risk. The percentage of patients with a complete response was higher in the transplantation group than in the RVD-alone group (59% vs. 48%, P=0.03), as was the percentage of patients in whom minimal residual disease was not detected (79% vs. 65%, P<0.001). Overall survival at 4 years did not differ significantly between the transplantation group and the RVD-alone group (81% and 82%, respectively). The rate of grade 3 or 4 neutropenia was significantly higher in the transplantation group than in the RVD-alone group (92% vs. 47%), as were the rates of grade 3 or 4 gastrointestinal disorders (28% vs. 7%) and infections (20% vs. 9%). No significant between-group differences were observed in the rates of treatment-related deaths, second primary cancers, thromboembolic events, and peripheral neuropathy. Among adults with multiple myeloma, RVD therapy plus transplantation was associated with significantly longer progression-free survival than RVD therapy alone, but overall survival did not differ significantly between the two approaches. (Supported by Celgene and others; IFM 2009 Study ClinicalTrials.gov number, NCT01191060 .).


Factors affecting the prevalence of infection with hepatitis B virus among non-pregnant women in the Alexishafen area of Papua New Guinea.
The prevalence of hepatitis B viral markers was studied in 673 women of childbearing age in 17 villages (12 indigenous and five plantation villages) on the north coast of Papua New Guinea. Some 7.9% of women were HBsAg positive and 41.3% were positive for anti-HBs. There was significant variation in prevalence between villages, ranging from 0 to 13.9% for HBsAg and 26.0 to 71.0% for all markers. The 12 indigenous villages were classified into three groups according to language (Austronesian or non-Austronesian), location (inland or coastal), and marriage patterns. The prevalence of hepatitis B was significantly higher in Austronesian than in non-Austronesian villages (P less than 0.01), and it remained significant after controlling for age differences and for possible effects on prevalence caused by women marrying into the three village groups from other areas. Interactions between malaria and hepatitis B were also investigated. Non-Austronesian villages with the highest spleen rates had the lowest prevalence of hepatitis B infection, and there was no correlation with parasitaemia. These results may reflect a lower exposure of women to hepatitis B infection in non-Austronesian villages, or may indicate different genetic or immunological responses to infection between Austronesians and non-Austronesians.


Aberrant axon initial segment plasticity and intrinsic excitability of ALS hiPSC motor neurons.
Dysregulated neuronal excitability is a hallmark of amyotrophic lateral sclerosis (ALS). We sought to investigate how functional changes to the axon initial segment (AIS), the site of action potential generation, could impact neuronal excitability in ALS human induced pluripotent stem cell (hiPSC) motor neurons. We find that early TDP-43 and C9orf72 hiPSC motor neurons show an increase in the length of the AIS and impaired activity-dependent AIS plasticity that is linked to abnormal homeostatic regulation of neuronal activity and intrinsic hyperexcitability. In turn, these hyperactive neurons drive increased spontaneous myofiber contractions of in vitro hiPSC motor units. In contrast, late hiPSC and postmortem ALS motor neurons show AIS shortening, and hiPSC motor neurons progress to hypoexcitability. At a molecular level, aberrant expression of the AIS master scaffolding protein ankyrin-G and AIS-specific voltage-gated sodium channels mirror these dynamic changes in AIS function and excitability. Our results point toward the AIS as an important site of dysfunction in ALS motor neurons.


Medicinal supplement genipin induces p53 and Bax-dependent apoptosis in colon cancer cells.
Colorectal cancer (CRC) is the third leading cause of cancer-associated mortality worldwide. Genipin is a medicinal herb compound derived from the gardenia fruit, which has been reported to exhibit antitumor activity against several types of cancer. The aim of the present study was to investigate the antitumor effect of genipin on colon cancer and the underlying molecular mechanisms. Genipin significantly inhibited the viability of HCT116 and SW480 cells in vitro in a dose- and time-dependent manner. Additionally, genipin was able to significantly inhibit tumor growth in nude mice with xenografts of HCT116 and SW480 cells. The inhibition of tumor growth by genipin treatment was coupled with G0/G1 cell cycle arrest, apoptosis induction, increased reactive oxygen species damage and loss of mitochondrial membrane potential. Further investigation of genipin-treated HCT116 cells revealed that the expression of p53, Bax and cleaved caspase-3 in genipin-treated cells was increased compared with the vehicle control, whereas B-cell lymphoma-2 expression appeared to be lower in genipin-treated cells. Collectively, the findings of the present study indicate that genipin was able to decrease proliferation and promote apoptosis in colon cancer cells by inducing the p53/Bax-mediated signaling pathway. Therefore, genipin may be used as a novel therapeutic agent in the treatment of CRC.


Evaluation of Psychiatric Symptomatology, Quality of Life, and Caregiver Burden in Mothers and Children with Primary Immunodeficiency.
The present study aimed to evaluate the quality of life, depression, and anxiety scores of children with primary immunodeficiency (PID) and depression, anxiety scores, and the caregiving burden of their mothers. A total of 149 children aged 2-18 years and their mothers were included in the present study, along with 125 healthy children and their mothers as a control group. The Pediatric Quality of Life Inventory (PedsQL), Child Depression Inventory (CDI), and Screening for Child Anxiety-Related Emotional Disorders (SCARED) questionnaire were used based on the views of children and their mothers. The Beck Depression Inventory (BDI), Beck Anxiety Inventory (BAI), Temperament Evaluation of Memphis, Pisa, Paris, San Diego Autoquestionnaire (TEMPS-A), and Zarit Caregiver Burden Scale (ZCB) were used for the mothers. According to children and their mothers, the scores of the PedsQL were lower than that of the control group (P < 0.05). In addition, according to the views of children and mothers, we found that PID children had higher depression and anxiety scores than healthy children (P < 0.05). The depression and anxiety levels of mothers in the patient group were also significantly higher than those in the control group (P = 0.05 and P = 0.001). Statistically, we found significantly lower psychosocial health summary scores and total scale score levels from the subclass of PedsQL in the patient group than in the control group. According to the views of both children and mothers, we observed that PID children had higher depression and anxiety scores than healthy children. It was also found that the BDI and BAI values in case of mothers in the patient group were significantly higher than those in the control group.


Study of Validity and Reliability of the Reinforcement Sensitivity Questionnaire in Turkish Community Sample.
In this study, it is aimed to assess the validity and reliability of Reinforcement Sensitivity Questionnaire (RSQ) which was developed on the basis of revised Reinforcement Sensitivity Theory (rRST) in Turkish community sample. The sample of study was consisted of 528 participants, 314 (59.5%) female and 214 (40.5%) male, in the range of 18-69 years. Data was collected by using online data collection tools via snowball method. Within the scope of the reliability study, test-retest application, split-half method and internal consistency analysis; within the scope of the validity study, explanatory factor analysis and confirmatory factor analysis were performed. For criterion-related validity Behavioral Inhibition System / Behavioral Activation System Scale (BIS/BAS), Eysenck Personality Questionnaire Revised-Abbreviated Form (EPQR-A) and State-Trait Anxiety Inventory (STAI-II) were applied. At the result of reliability analysis, the Cronbach Alpha internal consistency coefficient values of RSQ subscales were determined between 0.71 and 0.82. At the result of criterion-related validity, statistically significant correlations between subscales of RSQ and the similar subscales of BIS/BAS, EPQR-A and STAI-II were found. In explanatory factor analysis, a five-factor construct that has eigenvalues above 1, accounting for 49.96% of the total variance was found and it was seen that the questionnaire items loaded on their expected subscales consistent with the original questionnaire. In confirmatory factor analysis, CFI value was below the acceptable value of 0.90 at the first analysis. After eliminating two items and applying the recommended modifications, the fit indices reached the acceptable levels. According to the results of analysis, it was determined that Turkish version of RSQ is a valid and reliable evaluation tool for Turkish population.


Influence of nutrition on cognitive function in a group of elderly, independently living people.
To study the differences in cognitive status with respect to food habits and energy and nutrient intake in a group of non institutionalised, elderly people. Prospective study. The study subjects were independently living, elderly people who spent some of their time at day centres in the Comunidad de Madrid (the Madrid region). The study centres were selected by the Madrid City Hall. The study included 168 elderly people aged 65-90 y. All accepted the invitation to participate, met all inclusion criteria, and were free of significant cognitive impairment. Dietary intake was monitored using a 'food record' for 7 consecutive days including a Sunday. In addition, the 'precise individual weighing' method was used for 5 days in order to monitor the meals taken by the subjects at the centres' canteens. Cognitive capacity was measured using Folstein's mini-mental state examination (MMSE), validated for the Spanish population. Subjects with an adequate cognitive capacity (MMSE>/=28) showed a greater intake of total foods, fish, and alcoholic drinks, but took less foods from the 'various' group (chocolates, cakes, etc). These subjects had a more adequate intake of fatty acids and cholesterol, and a greater intake of vitamins implicated in correct brain function (thiamine, folic acid, vitamin C). Subjects with satisfactory intellectual function generally had a better diet. This shows the importance of correct nutrition in its maintenance.


Autoxidation of alpha-pinene at high oxygen pressure.
The liquid-phase oxidation of the renewable olefin alpha-pinene with molecular oxygen yields several valuable compounds for the fine-chemical industry. The most important products are verbenol/-one and alpha-pinene oxide. Following our previous work on the radical autoxidation at atmospheric pressure, this contribution addresses the influence of the oxygen pressure on the reaction mechanism and the product distribution. Trapping of the radical epoxide-precursor by O(2) causes a decrease of the epoxide selectivity, as well as the formation of a thermally unstable dialkylperoxide. This dialkylperoxide accelerates the rate significantly, due to an enhancement of the radical initiation. Although this causes a decrease of the radical chain-length, the amount of products produced in the chain-termination can still be neglected compared to the amount produced in the chain-propagations. Parallel to this, the ketone to alcohol ratio increases at higher oxygen pressure, due to the reaction of alkoxyl radicals with O(2), as well as a reaction of O(2) with the addition product of the alkoxyl radicals and the C=C double bond of the substrate. For O(2) partial pressures of 1 to 80 bar, rate constants of important reactions are extracted from the experimental observations via differential modelling, and confronted with literature values and/or quantum-chemical predictions. The derived mechanism is supported at the molecular level and provides a reliable description of the experimental observations.


Voluntary attention increases perceived spatial frequency.
Voluntary covert attention selects relevant sensory information for prioritized processing. The behavioral and neural consequences of such selection have been extensively documented, but its phenomenology has received little empirical investigation. Involuntary attention increases perceived spatial frequency (Gobell & Carrasco, 2005), but involuntary attention can differ from voluntary attention in its effects on performance in tasks mediated by spatial resolution (Yeshurun, Montagna, & Carrasco, 2008). Therefore, we ask whether voluntary attention affects the subjective appearance of spatial frequency--a fundamental dimension of visual perception underlying spatial resolution. We used a demanding rapid serial visual presentation task to direct voluntary attention and measured perceived spatial frequency at the attended and unattended locations. Attention increased the perceived spatial frequency of suprathreshold stimuli and also improved performance on a concurrent orientation discrimination task. In the control experiment, we ruled out response bias as an alternative account by using a lengthened interstimulus interval, which allows observers to disengage attention from the cued location. In contrast to the main experiment, the observers showed neither increased perceived spatial frequency nor improved orientation discrimination at the attended location. Thus, this study establishes that voluntary attention increases perceived spatial frequency. This phenomenological consequence links behavioral and neurophysiological studies on the effects of attention.


Malignant melanoma from the dermatologic perspective.
Early recognition and treatment of thin cutaneous melanomas (stages Ia and Ib) have contributed to a decreased case-fatality rate during the past several decades. The only known preventable risk factor for melanoma is sun protection in childhood, which directly affects the number of melanocytic nevi developing in an adult. Additional risk factors, clinical features, and the malignant potential of precursor lesions are discussed. The four major clinicopathologic subtypes of melanoma are described with recommendations for appropriate biopsy techniques for suspected melanoma. Nationwide skin cancer screenings by dermatologists and greater public awareness of the warning signs of melanoma have enhanced detection of early melanoma and have promoted chances for a cure.


Benzodiazepines: rat pinealocyte binding sites and augmentation of norepinephrine-stimulated N-acetyltransferase activity.
Studies of [3H]diazepam binding to intact rat pineal cells were carried out in tissue culture preparations. The binding was saturable, reversible and proportional to the number of cells used. Scatchard analysis resulted in a linear plot [Kd = 23 nM, maximum binding sites (Bmax) = 1.56 pmol/mg of protein for cells in monolayer culture; Kd = 7 nM, Bmax = 1.3 pmol/mg of protein for cells in suspension culture]. Inhibition constants (Ki) for clonazepam (500 nM), flunitrazepam (38 nM) and Ro-5-4864 (5 nM) indicated that the binding sites were probably of the "peripheral" type. In addition, the effects of diazepam on norepinephrine-stimulated N-acetyltransferase (NAT) activity were studied in organ culture and dissociated cell culture. Diazepam (10-50 microM) both prolonged and increased the magnitude of the norepinephrine-induced increase in NAT activity but did not affect the initial rate of rise of enzyme activity. The effect was dose-dependent and was also seen with clonazepam, flunitrazepam and Ro-5-4864, but not with Ro-15-1788. Diazepam, by itself, at these concentrations, had no effect on NAT, but enzyme activity was increased by higher concentrations (0.1-1 mM). Although a relationship between the [3H]diazepam binding sites described here and the effect of benzodiazepines on NAT cannot be established from these studies, the data suggest that the benzodiazepines may alter melatonin levels through their action on NAT.


Crossover versus mutation: a comparative analysis of the evolutionary strategy of genetic algorithms applied to combinatorial optimization problems.
Since their first formulation, genetic algorithms (GAs) have been one of the most widely used techniques to solve combinatorial optimization problems. The basic structure of the GAs is known by the scientific community, and thanks to their easy application and good performance, GAs are the focus of a lot of research works annually. Although throughout history there have been many studies analyzing various concepts of GAs, in the literature there are few studies that analyze objectively the influence of using blind crossover operators for combinatorial optimization problems. For this reason, in this paper a deep study on the influence of using them is conducted. The study is based on a comparison of nine techniques applied to four well-known combinatorial optimization problems. Six of the techniques are GAs with different configurations, and the remaining three are evolutionary algorithms that focus exclusively on the mutation process. Finally, to perform a reliable comparison of these results, a statistical study of them is made, performing the normal distribution z-test.


Monitoring defects on monolayer graphene using nematic liquid crystals.
Defects in graphene governs electrical and optical properties. Although grain boundaries in graphene inevitably formed during large area synthesis process, which act as scattering centers for charge carriers to degrade mobility, have been studied extensively, point defects have been rarely investigated mainly due to the absence of facile observation tools. Here, we report polarized optical microscopy to observe defect distributions in monolayer graphene. This was realized by aligning liquid crystal s (LC) on graphene where the defect population was modulated by irradiating ultraviolet (UV) light directly on graphene surface under moisture condition. Aromatic rings in LC molecules are oriented with hexagonal rings in graphene to have preferred orientation, providing a way to identify relative orientations of graphene domains and point defects. Our studies show that point defects generated by prolonged UV irradiation time give rise to irregular LC alignment with disclination lines on the graphene surface and a large-size LC domain associated with graphene single domain eventually disappeared. This indicates that defects associated with oxygen-containing functional groups cause to reduce the strong stacking interaction between graphene and LC molecules.


Alloreactive and syngeneic CTL are comparably dependent on interaction with MHC class I alpha-helical residues.
The molecular basis for the difference in the strength of T cell responses to self vs alloantigens is unknown, but may reflect how T cells are selected in the thymus. Because T cells with a high affinity for foreign as opposed to self MHC molecules are able to mature, it has been proposed that alloreactive T cells may be more strongly dependent upon interaction with MHC residues than are self-restricted T cells. This study was undertaken to rigorously address this hypothesis. Whereas other studies have compared self vs alloantigen recognition of different MHC alleles by a single T cell clone, we have compared self vs alloantigen recognition of a single MHC allele, H-2Ld, by a large panel of self-restricted and alloreactive T cell clones. Target cells expressing Ld molecules mutated at several different potential TCR contact residues were analyzed to determine which residues are important for recognition by self-restricted vs alloreactive T cells. We unequivocally demonstrate that self-restricted and alloreactive T cells do not differ, but rather are comparably dependent on interaction with MHC residues. Importantly, both self-restricted and alloreactive T cells are dependent upon the same MHC residues as primary contacts and, in addition, share a common recognition pattern of Ld. Furthermore, our analysis enables us to provide a model for allotype-specific T cell recognition of Ld vs Kb class I molecules.


The Naegleria genome: a free-living microbial eukaryote lends unique insights into core eukaryotic cell biology.
Naegleria gruberi, a free-living protist, has long been treasured as a model for basal body and flagellar assembly due to its ability to differentiate from crawling amoebae into swimming flagellates. The full genome sequence of Naegleria gruberi has recently been used to estimate gene families ancestral to all eukaryotes and to identify novel aspects of Naegleria biology, including likely facultative anaerobic metabolism, extensive signaling cascades, and evidence for sexuality. Distinctive features of the Naegleria genome and nuclear biology provide unique perspectives for comparative cell biology, including cell division, RNA processing and nucleolar assembly. We highlight here exciting new and novel aspects of Naegleria biology identified through genomic analysis.


Drought-induced stomatal closure probably cannot explain divergent white spruce growth in the Brooks Range, Alaska, USA.
Increment cores from the boreal forest have long been used to reconstruct past climates. However, in recent years, numerous studies have revealed a deterioration of the correlation between temperature and tree growth that is commonly referred to as divergence. In the Brooks Range of northern Alaska, USA, studies of white spruce (Picea glauca) revealed that trees in the west generally showed positive growth trends, while trees in the central and eastern Brooks Range showed mixed and negative trends during late 20th century warming. The growing season climate of the eastern Brooks Range is thought to be drier than the west. On this basis, divergent tree growth in the eastern Brooks Range has been attributed to drought stress. To investigate the hypothesis that drought-induced stomatal closure can explain divergence in the Brooks Range, we synthesized all of the Brooks Range white spruce data available in the International Tree Ring Data Bank (ITRDB) and collected increment cores from our primary sites in each of four watersheds along a west-to-east gradient near the Arctic treeline. For cores from our sites, we measured ring widths and calculated carbon isotope discrimination (δ13C), intrinsic water-use efficiency (iWUE), and needle intercellular CO2 concentration (C(i)) from δ13C in tree-ring alpha-cellulose. We hypothesized that trees exhibiting divergence would show a corresponding decline in δ13C, a decline in C(i), and a strong increase in iWUE. Consistent with the ITRDB data, trees at our western and central sites generally showed an increase in the strength of the temperature-growth correlation during late 20th century warming, while trees at our eastern site showed strong divergence. Divergent tree growth was not, however, associated with declining δ13C. Meanwhile, estimates of C(i) showed a strong increase at all of our study sites, indicating that more substrate was available for photosynthesis in the early 21st than in the early 20th century. Our results, which are corroborated by measurements of xylem sap flux density, needle gas exchange, and measurements of growth and δ13C along moisture gradients within each watershed, suggest that drought-induced stomatal closure is probably not the cause of 20th century divergence in the Brooks Range.


Statistical inferences for polarity identification in natural language.
Information forms the basis for all human behavior, including the ubiquitous decision-making that people constantly perform in their every day lives. It is thus the mission of researchers to understand how humans process information to reach decisions. In order to facilitate this task, this work proposes LASSO regularization as a statistical tool to extract decisive words from textual content in order to study the reception of granular expressions in natural language. This differs from the usual use of the LASSO as a predictive model and, instead, yields highly interpretable statistical inferences between the occurrences of words and an outcome variable. Accordingly, the method suggests direct implications for the social sciences: it serves as a statistical procedure for generating domain-specific dictionaries as opposed to frequently employed heuristics. In addition, researchers can now identify text segments and word choices that are statistically decisive to authors or readers and, based on this knowledge, test hypotheses from behavioral research.


Immunocytochemistry of cell surface heparan sulfate proteoglycan in mouse tissues. A light and electron microscopic study.
The core protein of the proteoglycan at the cell surface of NMuMG mouse mammary epithelial cells bears both heparan and chondroitin sulfate chains and is recognized by the monoclonal antibody 281-2. Using this antibody and the peroxidase-antiperoxidase staining technique in adult mouse tissues, we found that the antibody recognizes the antigen in a highly restricted distribution, staining a variety of epithelial cells but no cells derived from embryonic mesoderm or neural crest. The antibody fails to stain any stromal (mesenchymal) or neuronal cells, with the exception of plasma cells and Leydig cells. Squamous and transitional epithelia stain intensely over their entire surfaces, whereas cuboidal and columnar epithelia stain moderately and only at the lateral surface of the basal cells. Within squamous and transitional epithelial tissues that undergo physiological regeneration (e.g., epidermis), the most superficial and differentiated cell types fail to stain. Within glandular and branched epithelia (e.g., pancreas), the secretory alveolar cells fail to stain. When evaluated by electron microscopy, granular deposits of stain are seen on the plasma membrane, especially on lateral surfaces, but none are noted within the cells or the basement membrane. These results indicate that in adult tissues the core protein of this heparan sulfate-rich proteoglycan is expressed almost exclusively at epithelial cell surfaces. Expression appears to be lost as the cells become either mature or highly differentiated.


Qualitative Behavioural Assessment as a welfare indicator for farmed Atlantic salmon (Salmo salar) in response to a stressful challenge.
Animal welfare assessments have struggled to investigate the emotional states of animals while focusing solely on available empirical evidence. Qualitative Behavioural Assessment (QBA) may provide insights into an animal's subjective experiences without compromising scientific rigor. Rather than assessing explicit, physical behaviours (i.e., what animals are doing, such as swimming or feeding), QBA describes and quantifies the overall expressive manner in which animals execute those behaviours (i.e., how relaxed or agitated they appear). While QBA has been successfully applied to scientific welfare assessments in a variety of species, its application within aquaculture remains largely unexplored. This study aimed to assess QBA's effectiveness in capturing changes in the emotional behaviour of Atlantic salmon following exposure to a stressful challenge. Nine tanks of juvenile Atlantic salmon were video-recorded every morning for 15 min over a 7-day period, in the middle of which a stressful challenge (intrusive sampling) was conducted on the salmon. The resultant 1-min, 63 video clips were then semi-randomised to avoid predictability and treatment bias for QBA scorers. Twelve salmon-industry professionals generated a list of 16 qualitative descriptors (e.g., relaxed, agitated, stressed) after viewing unrelated video-recordings depicting varying expressive characteristics of salmon in different contexts. A different group of 5 observers, with varied experience of salmon farming, subsequently scored the 16 descriptors for each clip using a Visual Analogue Scale (VAS). Principal Components Analysis (correlation matrix, no rotation) was used to identify perceived patterns of expressive characteristics across the video-clips, which revealed 4 dimensions explaining 74.5% of the variation between clips. PC1, ranging from 'relaxed/content/positive active' to 'unsettled/stressed/spooked/skittish' explained the highest percentage of variation (37%). QBA scores for video-clips on PC1, PC2, and PC4 achieved good inter- and intra-observer reliability. Linear Mixed Effects Models, controlled for observer variation in PC1 scores, showed a significant difference between PC1 scores before and after sampling (p = 0.03), with salmon being perceived as more stressed afterwards. PC1 scores also correlated positively with darting behaviours (r = 0.42, p < 0.001). These results are the first to report QBA's sensitivity to changes in expressive characteristics of salmon following a putatively stressful challenge, demonstrating QBA's potential as a welfare indicator within aquaculture.


Comparison of the T cell patterns in leprous and cutaneous sarcoid granulomas. Presence of Valpha24-invariant natural killer T cells in T-cell-reactive leprosy together with a highly biased T cell receptor Valpha repertoire.
The T-cell-reactive (eg, tuberculoid and reversal) forms of leprosy represent a well-defined granulomatous reaction pattern against an invading pathogen. The immune response in cutaneous sarcoidosis is a granulomatous condition that pathologically is very similar to T-cell reactive leprosy. However, it lacks a defined causative agent. In view of the role of NKT cells in murine granulomas induced by mycobacterial cell walls, we have searched for the presence of NKT cells in the cutaneous lesions of both leprosy and sarcoidosis. These cells were present in T-cell-reactive leprosy but were undetectable in cutaneous sarcoidosis. We have also studied the TCR Valpha repertoire in the two diseases. In addition to Valpha24(+) NKT cells, all patients with T-cell-reactive leprosy showed a very restricted T-cell-reactive Valpha repertoire with a strong bias toward the use of the Valpha6 and Valpha14 segments. Valpha6 and Valpha14(+) T cells were polyclonal in terms of CDR3 length and Jalpha usage. In contrast, most sarcoidosis patients showed a diverse usage of Valpha chains associated with clonal or oligoclonal expansions reminiscent of antigen-driven activation of conventional T cells. Thus the origin and perpetuation of the two kinds of granulomatous lesions appear to depend on altogether distinct T-cell recruiting mechanisms.


Calmodulin antagonists depress calcium and potassium currents in ventricular and vascular myocytes.
Myocytes isolated from guinea pig ventricles or bovine portal veins were voltage clamped with a single patch electrode. The calmodulin antagonists (CaM-A) calmidazolium, trifluoperazine (TFP), and chlorpromazine acted as Ca antagonists; they reduced the calcium inward current ICa in a voltage- and use-dependent way. For ventricular myocytes, 50% effective concentration (EC50) of calmidazolium was 1 microM, and the EC50 for TFP was 2.5 microM. For vascular myocytes, these numbers were 0.3 and 1 microM, respectively. CaM-A moderately retarded the inactivation time course and shifted the ICa availability curve to more negative potentials. CaM-A were not selective Ca antagonists; other membrane currents such as sodium currents and inwardly and delayed potassium currents were reduced as well (EC50 between 5 and 10 microM). It is unlikely that the above effects require binding of CaM-A to Ca-calmodulin, since reduction of ICa or potassium current (IK) was not modified when 1) the cells were loaded with 100 microM exogenous calmodulin or 2) Ca ions were removed from the extra- and intracellular space. Instead, the unspecific reduction of membrane currents may result from a change in the lipids of the sarcolemma into which CaM-A partition and accumulate.


Age at onset in genetic prion disease and the design of preventive clinical trials.
To determine whether preventive trials in genetic prion disease could be designed to follow presymptomatic mutation carriers to onset of disease. We assembled age at onset or death data from 1,094 individuals with high penetrance mutations in the prion protein gene (PRNP) in order to generate survival and hazard curves and test for genetic modifiers of age at onset. We used formulae and simulations to estimate statistical power for clinical trials. Genetic prion disease age at onset varies over several decades for the most common mutations and neither sex, parent's age at onset, nor PRNP codon 129 genotype provided additional explanatory power to stratify trials. Randomized preventive trials would require hundreds or thousands of at-risk individuals in order to be statistically powered for an endpoint of clinical onset, posing prohibitive cost and delay and likely exceeding the number of individuals available for such trials. The characterization of biomarkers suitable to serve as surrogate endpoints will be essential for the prevention of genetic prion disease. Parameters such as longer trial duration, increased enrollment, and the use of historical controls in a postmarketing study could provide opportunities for subsequent determination of clinical benefit.


Spatial Control of Microbial Pesticide Degradation in Soil: A Model-Based Scenario Analysis.
Microbial pesticide degraders are heterogeneously distributed in soil. Their spatial aggregation at the millimeter scale reduces the frequency of degrader-pesticide encounter and can introduce transport limitations to pesticide degradation. We simulated reactive pesticide transport in soil to investigate the fate of the widely used herbicide 4-chloro-2-methylphenoxyacetic acid (MCPA) in response to differently aggregated distributions of degrading microbes. Four scenarios were defined covering millimeter scale heterogeneity from homogeneous (pseudo-1D) to extremely heterogeneous degrader distributions and two precipitation scenarios with either continuous light rain or heavy rain events. Leaching from subsoils did not occur in any scenario. Within the topsoil, increasing spatial heterogeneity of microbial degraders reduced macroscopic degradation rates, increased MCPA leaching, and prolonged the persistence of residual MCPA. In heterogeneous scenarios, pesticide degradation was limited by the spatial separation of degrader and pesticide, which was quantified by the spatial covariance between MCPA and degraders. Heavy rain events temporarily lifted these transport constraints in heterogeneous scenarios and increased degradation rates. Our results indicate that the mild millimeter scale spatial heterogeneity of degraders typical for arable topsoil will have negligible consequences for the fate of MCPA, but strong clustering of degraders can delay pesticide degradation.


Physicochemical properties determining drug detection in skin.
Chemicals, including some systemically administered xenobiotics and their biotransformations, can be detected noninvasively using skin swabs and untargeted metabolomics analysis. We sought to understand the principal drivers that determine whether a drug taken orally or systemically is likely to be observed on the epidermis by using a random forest classifier to predict which drugs would be detected on the skin. A variety of molecular descriptors describing calculated properties of drugs, such as measures of volume, electronegativity, bond energy, and electrotopology, were used to train the classifier. The mean area under the receiver operating characteristic curve was 0.71 for predicting drug detection on the epidermis, and the SHapley Additive exPlanations (SHAP) model interpretation technique was used to determine the most relevant molecular descriptors. Based on the analysis of 2561 US Food and Drug Administration (FDA)-approved drugs, we predict that therapeutic drug classes, such as nervous system drugs, are more likely to be detected on the skin. Detecting drugs and other chemicals noninvasively on the skin using untargeted metabolomics could be a useful clinical advancement in therapeutic drug monitoring, adherence, and health status.


Minimally Invasive Circumferential Hiatal Dissection for the Treatment of Adenocarcinoma of the Distal Esophagus and Esophago-gastric Junction: Technical Considerations Combined With Histopathological Outcomes.
Circumferential resection margin involvement is an independent prognostic factor in patients with adenocarcinoma of the distal esophagus and esophago-gastric junction. However, there is currently no consensus on the extent and the technique of hiatal dissection. We describe a minimally invasive technique of circumferential hiatal dissection for adenocarcinoma of the distal esophagus and esophago-gastric junction with its related histopathological results. A prospective study of 40 consecutive patients undergoing hybrid (laparoscopic/thoracotomic) or totally minimally invasive Ivor-Lewis esophagogastrectomy over a period of 21 months was conducted. Dissection of the hiatus included peri-esophageal surrounding tissues in a cylindrical fashion maximizing the distance from the oesophageal wall. Crural muscle fibers and pleura bilaterally, pericardial fat anteriorly and pre-aortic tissue posteriorly were excised en bloc. Histopathological results particularly focused on involvement of the circumferential resection margin. Neoadjuvant chemotherapy was given to 24 (60%) patients. Complete histological clearance (R0) was achieved in 92.5% (n=37) according to the criteria of the College of American Pathologists and in 87.5% (n=35) according to those of the Royal College of Pathologists. In pT3 tumors (n=22), the circumferential resection margin was negative in 20 patients (91%) according to the College of American Pathologists, and in 17 (77%) according to the Royal College of Pathologists. Adoption of this safe and reproducible technique might reduce the incidence of circumferential resection margin involvement and improve pathological outcomes. In addition, there may be positive implications for training and quality control.


[Clinical picture and characteristics of the course of tic hyperkinesis in children].
The author examined 90 children with ticose hyperkinesis. On the basis of a thorough study of the clinical picture, course of the disease and the use of laboratory methods all the patients were divided into 3 groups: with regressive, remittent and progradient development and characteristic clinical signs. For each group a corresponding treatment with a good effect was selected.


Effects of in vitro-growth culture duration on fertilizability of bovine growing oocytes and proliferation of cells surrounding oocytes.
The effects of in vitro-growth (IVG) culture duration (12 and 14 days) on the fertilizability and the developmental ability of IVG oocytes matured with pre-maturational culture, and on the proliferation of cells surrounding oocytes during IVG culture were examined. The fertilization and cleavage rates of 12- and 14-day IVG oocytes were similar; however, 14-day IVG oocytes showed lower blastocyst development rate than 12-day IVG oocytes. In addition, the number of cells surrounding oocytes increased until 12 days of IVG, but decreased at 14 days. The results indicated that the extension of IVG period beyond 12 days impaired the proliferation of cells surrounding oocytes, leading to the decrease of developmental ability but not fertilizability of 14-day IVG oocytes.


Epidemiological profile of malaria in a university hospital in the eastern region of Saudi Arabia.
To study the epidemiological, clinical and hematological profile of laboratory-diagnosed malaria cases at King Fahd Hospital of the University, Al-Khobar, Saudi Arabia, during the period from January 1990 to December 1999, and to provide suitable recommendations accordingly. This was a clinical case series study of confirmed cases presenting to King Fahd Hospital of the University during the period from January 1990 to December 1999. A specially designed form was used for data collection and 602, laboratory-confirmed cases of malaria were retrospectively analyzed. There were 602 cases with a mean age of 25.8 + 14.3 and a male to female ratio of 2.9:1. Less than half the cases were Saudis (42%), most of whom (93%) reported a history of travel to the Southwestern part of the Kingdom. The highest frequency of cases was observed in the years 1992, 1994 and 1998 and 40% of the cases were diagnosed during the months of February, March and September. Plasmodium falciparum was the most common species among Saudi (83%), Sudanese (72%) and Yemeni (64%) patients, while Plasmodium vivax was predominant among others. Most of these cases (75%) had a history of travel to their home countries (endemic areas). The most common clinical presentation was fever (97%), while the most common clinical signs were splenomegaly (9%) and jaundice (8%). Anemia (60%) and thrombocytopenia (53%) were the most common hematological findings. Although it appears that the Eastern Province is still free of indigenous malaria transmission, this could not be confirmed by the data. Imported cases, however represent a continuous threat due to the existence of such vectors as Anopheles stephensi, Anopheles fluviatilis, Anopheles sergentii and Anopheles superpictus and a large number of non-immune persons. It is recommended that malaria be always considered in the differential diagnosis of all acute fevers, especially among those with a history of travel to an endemic area. Prompt diagnosis and treatment is necessary. Chemoprophylaxis, when traveling to endemic areas is mandatory, as well as the use of other primary preventive measures to protect against mosquito bites.


Haemoglobin potentiates the respiration-inhibitory action of lipoxygenases via its pseudolipohydroperoxidase activity.
The inhibition of the NADH oxidase and of the succinate-cytochrome c oxidoreductase activities of beef heart submitochondrial particles (ETP) induced by reticulocyte lipoxygenase is strongly potentiated by haemoglobin added simultaneously or subsequently to lipoxygenase. Half-maximal potentiation was observed with 0.9 microM haemoglobin. Haemoglobin augments also the lipoxygenase-induced loss of acid-labile sulphur in ETP not related to respiratory inhibition. Soybean lipoxygenase produces only weak respiratory inhibition even at high concentrations in the absence of haemoglobin; strong inhibition is obtained, however, in the presence of haemoglobin. Among other haem compounds tested only free haemin but not cytochrome c and catalase exhibit identical effects. The action of haem compounds on the system ETP plus lipoxygenase is accompanied by their destruction. The experimental evidence indicates that the effects of haemoglobin described here are due to its ability to decompose catalytically hydroperoxy lipids generated by lipoxygenase. This catalytic property (pseudolipohydroperoxidase activity) was demonstrated with 13L-hydroperoxylinoleic acid as substrate. The relations between the various catalytic actions of haemoglobin related to lipid peroxidation are discussed.


Termination-reinitiation occurs in the translation of mammalian cell mRNAs.
Many examples of internal translation initiation in eucaryotes have accumulated in recent years. In many cases terminators of upstream reading frames precede the internal initiation site, suggesting that translational reinitiation may be a mechanism for initiation at internal AUGs. To test this idea, a series of recombinants was constructed in the mammalian expression vector pSV2. Each contained a dicistronic transcription unit comprising the coding sequence for mouse dihydrofolate reductase (DHFR) followed by the gene for xanthine-guanine phosphoribosyl transferase (XGPRT) from Escherichia coli. Various versions of this pSV2dhfr-gpt recombinant plasmid altered the location at which the DHFR reading frame was terminated relative to the XGPRT initiation codon and demonstrated that this is a critical factor for the expression of XGPRT activity in transfected Cos-1 cells. Thus, when the DHFR frame terminated upstream or a very short distance downstream of the XGPRT initiator AUG, substantial levels of XGPRT activity were observed. When the DHFR frame terminated 50 nucleotides beyond the XGPRT initiator, activity was reduced about twofold. However, when the DHFR and XGPRT sequences were fused in-frame so that ribosomes which initiated at the DHFR AUG did not terminate until they encountered the XGPRT terminator, production of XGPRT activity was abolished. This dependence of internal translation initiation on the position of terminators of the upstream reading frame is consistent with the hypothesis that mammalian ribosomes are capable of translational reinitiation.


Two new species of Tetragonocephalum (Cestoda: Lecanicephalidea) from Pastinachus sephen (Myliobatiformes: Dasyatidae) from the Gulf of Oman.
In the present study two new species of Tetragonocephalum Shipley et Hornell, 1905, T. mackenziei sp. n. and T. kazemii sp. n., are described from the spiral intestine of the cowtail stingray, Pastinachus sephen (Forsskål), from the northern coast of the Gulf of Oman. Tetragonocephalum mackenziei is distinguished from the 16 other valid species of Tetragonocephalum by a unique combination of characteristics, i.e. sperm-filled seminal receptacle in immature proglottids, body length (7.7-17.5 mm), body width (213-288 µm), number of proglottids (34-49), number of testes (10-14), size of scolex (228-315 µm × 213-288 µm) and size of acetabula (56-73 µm × 61-75 µm). Tetragonocephalum kazemii is morphologically distinguishable from its valid congeners and T. mackenziei based on a combination of characteristics, including body length (28.8-36.6 mm), number of proglottids (50-65), number of testes (30-42), size of scolex (388-564 µm × 326-448 µm), size of acetabula (62-86 µm × 57-90 µm) and testes (25-39 × 21-32). This brings the total number of validly described species of Tetragonocephalum to 18 and expands our knowledge of this diverse genus to now include the Gulf of Oman, as well as Arafura Sea, northern Indian Ocean and western Pacific Ocean.


Neonatal brain abnormalities and memory and learning outcomes at 7 years in children born very preterm.
Using prospective longitudinal data from 198 very preterm and 70 full term children, this study characterised the memory and learning abilities of very preterm children at 7 years of age in both verbal and visual domains. The relationship between the extent of brain abnormalities on neonatal magnetic resonance imaging (MRI) and memory and learning outcomes at 7 years of age in very preterm children was also investigated. Neonatal MRI scans were qualitatively assessed for global, white-matter, cortical grey-matter, deep grey-matter, and cerebellar abnormalities. Very preterm children performed less well on measures of immediate memory, working memory, long-term memory, and learning compared with term-born controls. Neonatal brain abnormalities, and in particular deep grey-matter abnormality, were associated with poorer memory and learning performance at 7 years in very preterm children. Findings support the importance of cerebral neonatal pathology for predicting later memory and learning function.


Metals and cellular signaling in mammalian cells.
A number of heavy metals are known to be essential for life, but most of these can also be toxic to cells under certain circumstances, or at elevated levels. Metals can directly induce gene expression through the actions of metal-responsive transcription factors. However, metals can also influence the response to non-metal extracellular signals. Cells respond to extracellular signals through a variety of different, but often interacting, signal transduction pathways. Metals can alter cell behaviour by interacting with transcription factors and transduction molecules, many of which are dependent on metals (primarily zinc) for their action. In addition, metals can affect cells in more nonspecific ways, for example, by inducing a generalized stress response or by cross-linking cell surface thiol groups. The prominent role of zinc in signal transduction combined with low intracellular free zinc levels has lead to the speculation that cellular signaling and gene expression may be regulated, in part, by zinc bioavailability. Experimental modification of the levels of the intracellular metal-binding protein, metallothionein (MT), results in altered responsiveness to extracellular signals. This observation suggests that MT is capable of influencing gene expression, perhaps by regulating the level of intracellular free zinc.


Significance of headache in intracranial vertebrobasilar artery dissections: an observational study.
Headache may represent acute phase of intracranial vertebrobasilar artery dissection (iVBAD). We aimed to evaluate its clinical significance in iVBAD. Consecutive acute iVBAD patients were grouped into ruptured iVBAD, unruptured iVBAD with no headache, isolated headache, or concurrent headache with neurological symptoms. Composite hemorrhagic/ischemic endpoints, and dynamic arterial changes were graded. Clinical characteristics of the four groups, and association between headache and composite outcomes was evaluated. Headaches were precedent in 79% of the ruptured iVBAD patients (maximal delay, 10D). In unruptured iVBAD, when patients with no headache (N = 69), concurrent headache (N = 111), and isolated headache (N = 126) were compared, concurrent headache was associated with ischemic endpoints (isolated headache as reference, adjusted odds ratio: 6.40, 95% confidence interval [2.03-20.19]). While there were no differences in hemorrhagic endpoints, dynamic arterial changes were higher in the isolated headache group (aOR: 3.98, 95% CI [1.72-9.18]) but not for the concurrent headache group (aOR: 1.59 [0.75-3.38]) compared to no headache group. Headache was more commonly severe (48.4% vs. 17.3%, p < 0.001) and ipsilateral (59.7% vs. 45.5%, p = 0.03) for isolated headache compared to concurrent headache, indicating a higher causal relationship. In iVBAD, isolated headache may be considered an acute-phase biomarker, associated with dynamic arterial changes.


Cold Adaptation of Triosephosphate Isomerase.
The main problem for enzymes from psychrophilic species, which need to work near the freezing point of liquid water, is the exponential decay of reaction rates as the temperature is decreased. Cold-adapted enzymes have solved this problem by shifting the activation enthalpy-entropy balance for the catalyzed reaction compared to those of their mesophilic orthologs. To understand the structural basis of this universal feature, it is necessary to examine pairs of such orthologous enzymes, with known three-dimensional structures, at the microscopic level. Here, we use molecular dynamics free energy calculations in combination with the empirical valence bond method to evaluate the temperature dependence of the activation free energy for differently adapted triosephosphate isomerases. The results show that the enzyme from the psychrophilic bacterium Vibrio marinus indeed displays the characteristic shift in enthalpy-entropy balance, compared to that of the yeast ortholog. The origin of this effect is found to be located in a few surface-exposed protein loops that show differential mobilities in the two enzymes. Key mutations render these loops more mobile in the cold-adapted triosephosphate isomerase, which explains both the reduced activation enthalpy contribution from the protein surface and the lower thermostability.


Malignant neoplasms of respiratory and intrathoracic organs (C30-C39) in the Osijek-Baranja County, Croatia.
The Institute of Public Health of the Osijek-Baranja County in collaborate with different county institutes provide updated information on the cancer occurrence and trends in the Osijek-Baranja County (OBC). The cancers were defined according to the International Statistical Classification of Diseases and Related Health Problems, 10th Revision (ICD-10), codes of malignant neoplasms of respiratory and intrathoracic organs (C30-C39). The aim of this article was to show the size of cancer problem with the C30-C39 cancer group in the Osijek-Baranja County (OBC). This article processes data on cancer incidence and mortality, appertaining age distribution, median age, cancer survival and length of stay in hospital collected in period 2001-2009. Out of all patients diagnosed with C30-C39 cancers, there were 18.2% of females and 81.8% of males. The total incidence rate in males (119.5/100,000) decreases while the total mortality rate (110.9/100,000) does not change in 9-year period. In the same period, the total mortality rate in females (15.7/100,000) increase moderately. The age-standardized incidence rate was six times higher in males than in females. The overall median age at diagnosis of C30-C39 cancers of both genders was 64.5 years, which exceeds the average age at diagnosis of cancer in general in the OBC by 4.8 years. Five-year relative survival rate was 14.8%, 19.7% for females and 13.7% for males. Male lung and bronchus cancer patients (C34) were 1 year younger at diagnosis of cancer than the respective female patients. An average C30-C39 cancer patient was hospitalized 2.0 times during the course of their illness while the median length of stay in hospital amounted to 16.1 days. The number of hospital admissions in both genders decreased over the 2001-2009 period. In both genders, the total length of stay in hospitals was slightly reduced. Females spent 0.4 days more in hospital than males. The overall incidence and mortality rate in the OBC were among the highest in Europe. However, these rates in females reached neither the Croatian nor the EU average. Other data are similar to those in Europe.


Probing the influence of cell surface polysaccharides on nanodendrimer binding to Gram-negative and Gram-positive bacteria using single-nanoparticle force spectroscopy.
The safe use and design of nanoparticles (NPs) ask for a comprehensive interpretation of their potentially adverse effects on (micro)organisms. In this respect, the prior assessment of the interactions experienced by NPs in the vicinity of - and in contact with - complex biological surfaces is mandatory. It requires the development of suitable techniques for deciphering the processes that govern nano-bio interactions when a single organism is exposed to an extremely low dose of NPs. Here, we used atomic force spectroscopy (AFM)-based force measurements to investigate at the nanoscale the interactions between carboxylate-terminated polyamidoamine (PAMAM) nanodendrimers (radius ca. 4.5 nm) and two bacteria with very distinct surface properties, Escherichia coli and Lactococcus lactis. The zwitterionic nanodendrimers exhibit a negative peripheral surface charge and/or a positive intraparticulate core depending on the solution pH and salt concentration. Following an original strategy according to which a single dendrimer NP is grafted at the very apex of the AFM tip, the density and localization of NP binding sites are probed at the surface of E. coli and L. lactis mutants expressing different cell surface structures (presence/absence of the O-antigen of the lipopolysaccharides (LPS) or of a polysaccharide pellicle). In line with electrokinetic analysis, AFM force measurements evidence that adhesion of NPs onto pellicle-decorated L. lactis is governed by their underlying electrostatic interactions as controlled by the pH-dependent charge of the peripheral and internal NP components, and the negatively-charged cell surface. In contrast, the presence of the O-antigen on E. coli systematically suppresses the adhesion of nanodendrimers onto cells, may the apparent NP surface charge be determined by the peripheral carboxylate groups or by the internal amine functions. Altogether, this work highlights the differentiated roles played by surface polysaccharides in mediating NP attachment to Gram-positive and Gram-negative bacteria. It further demonstrates that the assessment of NP bioadhesion features requires a critical analysis of the electrostatic contributions stemming from the various structures composing the stratified cell envelope, and those originating from the bulk and surface NP components. The joint use of electrokinetics and AFM provides a valuable option for rapidly addressing the binding propensity of NPs to microorganisms, as urgently needed in NP risk assessments.


The Effect of Vitamin B12 Infusion on Prevention of Nitrous Oxide-induced Homocysteine Increase: A Double-blind Randomized Controlled Trial.
Nitrous oxide is a common inhalation anesthetic agent in general anesthesia. While it is widely accepted as a safe anesthetic agent, evidence suggests exposure to this gas, leads to hyperhomocysteinemia. The present study aimed to evaluate the effects of single-dose intravenous infusions of vitamin B12, before and after the induction of nitrous oxide anesthesia on homocysteine levels after the surgery. This double-blind randomized controlled trial was conducted on 60 patients who were scheduled for elective surgery under general anesthesia, presumably lasting for more than two hours. The subjects were randomly allocated to three groups of 20. For the first group, vitamin B12 solution (1 mg/100 ml normal saline) and 100 ml of normal saline (placebo), were infused before and after the induction of anesthesia, respectively. The second group received placebo and vitamin B12 infusion before and after the induction of anesthesia, respectively. The third group received placebo infusions at both times. Homocysteine levels were measured before and 24 hours after the surgery. The mean homocysteine and vitamin B12 levels were significantly different within the three groups (p<0.001). In patients who had been infused with vitamin B12 before the surgery, homocysteine levels were significantly lower than the other two groups. In the placebo group, homocysteine levels significantly increased after the surgery. Nitrous oxide causes hyperhomocysteinemia after general anesthesia. Since vitamin B12 infusion is a safe and inexpensive method to decrease homocysteine levels in these patients, it may be recommended for patients undergoing nitrous oxide anesthesia to be used before induction of anesthesia.


Electronic structure calculations as a tool for investigating acyl migrations in ester saponins.
The possibility of acyl migrations in ester saponins from Maesa lanceolata was investigated by molecular mechanics and electronic structure calculations carried out on the major constituent maesasaponin IV3 (3beta-O-[[alpha-L-rhamnopyranosyl-(1-->2)-[beta-D-galactopyranosyl-(1-->3)]-[beta-D-galactopyranosyl-(1-->2)]-beta-D-gluco-pyranuronyl]-21beta-angeloyloxy-22alpha-propanoyloxy-13beta,28-oxido-olean-16alpha, 28alpha-diol). It was confirmed that acyl migrations could occur in rings D and E of maesasaponins.


Cancer chemoprevention with PV-1, a novel Prunella vulgaris-containing herbal mixture that remodels the tumor immune microenvironment in mice.
The herb Prunella vulgaris has shown significant immune-stimulatory and anti-inflammatory effects in mouse models. Here, the effects of a novel Prunella vulgaris-containing herbal mixture, PV-1, were examined in several mouse models for cancer, including chemically induced models of lung and oral cancers as well as syngraft models for lung cancer and melanoma. PV-1, consisting of extracts from Prunella vulgaris, Polygonum bistorta, Sonchus brachyotus and Dictamnus dasycarpus, exhibited no toxicity in a dose escalation study in A/J mice. PV-1 significantly inhibited mouse lung tumor development induced by the lung carcinogens vinyl carbamate and benzo[a]pyrene. PV-1 also hindered the induction of oral squamous cell carcinomas in C57BL/6 mice caused by 4-nitroquinoline-1-oxide. Flow cytometry analysis showed that PV-1 increased the numbers of CD8+ tumor-infiltrating lymphocytes (TILs) and increased the production of granzyme B, TNF-α, and IFN-γ by CD8+ TILs. PV-1 also suppressed granulocytic myeloid-derived suppressor cell numbers (g-MDSCs) and improved the anti-cancer activity of anti-PD-1 immunotherapy. These results indicate that PV-1 remodels the tumor immune microenvironment by selectively inhibiting g-MDSCs and increasing CD8+ TILs within tumors, resulting in decreased immune suppression and enhanced cancer chemopreventive efficacy.


Biomechanical consequences of first metatarsal osteotomy in treating hallux valgus.
Among the numerous osteotomies for correction of hallux valgus, the modified chevron is known for its good intrinsic stability and the scarf for its large corrective potential. An intermediate design, the reversed-L osteotomy, has been developed to combine these competing biomechanical objectives. The purpose of this in vitro study was to compare the structural and local biomechanical performance of these three designs. Stiffness, cortical bone strains (a factor relevant to bone remodeling), strength and failure mode of the scarf, modified chevron and reversed-L osteotomies were measured on human specimens in two different loading configurations. The scarf osteotomy caused significant changes in stiffness and cortical bone strains with the proximal apex being at the origin of bone failure. The chevron and reversed-L had a generally comparable response to the intact bone. The chevron specimens failed by pivoting of the distal fragment, and the reversed-L by pivoting or fracture. This is the first study to investigate the cortical bone strain changes induced by these invasive osteotomies. Alterations from the intact bone response could be directly related to the design of the osteotomy. Notably, the critical weakening proximal apex of the scarf is avoided in the reversed-L, leading to results comparable to the chevron. This study provides support in favor of the intermediate design of the reversed-L as an effective compromise between the competing biomechanical objectives of corrective potential and mechanical stability.


Organization and expression of the immediate early genes of human cytomegalovirus.
The immediate early genes of human cytomegalovirus were characterized according to map location, RNA transcripts, and translation products. Three regions in the large unique component (0.709 to 0.751 map units) were transcribed in the presence of an inhibitor of protein synthesis (anisomycin). A single size class of polyadenylated mRNA, 1.95 kilobases (kb), was transcribed abundantly relative to the other size classes. The predominant 1.95-kb viral RNA was transcribed from right to left on the prototype arrangement of the viral genome and spanned a region of approximately 2.8 kb (0.739 to 0.751 map units). This mRNA codes for a 75,000-dalton protein that represents the predominant immediate early protein detected in infected cells. Immunoprecipitation of viral proteins synthesized in vitro as well as in vivo demonstrated that the predominant immediate early protein is synthesized as a protein of 75,000 daltons, but is presumably modified in vivo, resulting in a broad banding pattern ranging from 75,000 to 68,000 daltons. A different immediate early viral gene (0.732 to 0.739 map units) is transcribed from left to right at relatively low levels. The 3' ends of the above viral RNAs terminate at approximately 230 base pairs apart in the region of approximately 0.739 map units. Five RNA size classes ranging from 2.25 to 1.10 kb were detected, but the 1.75-kb and 1.40-kb RNA size classes were more abundant from this region. At least four minor proteins are coded by these mRNAs, with apparent molecular weights ranging from 56,000 to 16,500. Last, a 1.95-kb mRNA was transcribed from a third region (0.709 to 0.728 map units). This viral mRNA was present at relatively low concentration and coded for a minor protein of 68,000 daltons. Since immediate early gene expression of human cytomegalovirus is dominated by the synthesis of an mRNA from the region of 0.739 to 0.751 map units that codes for the predominant immediate early protein found in the infected cell, we hypothesize that this protein is the major regulatory protein influencing the switch from restricted to extensive transcription.


Comprehensive Women-centered Treatment for Substance Use Disorders in Georgia: Current Status and Future Directions.
The present paper examines the current status of women-centered substance use disorder treatment in Georgia. Four major issues are identified that adversely impact the delivery of effective services for women with substance use disorders: Policy Issues; Sociocultural Issues; Programmatic/Structural Issues; and Personal/Interpersonal Issues. These four issues are seen to form a complex, dynamic system that serves to maintain the current ineffective service delivery system and suppresses movement toward an effective service delivery for this highly marginalized and at-risk population. How these issues, and their interplay, present continuing barriers to the development and implementation of effective treatment for this population are outlined and discussed. In order to overcome these barriers, solutions must be sought in four areas: Policy reform; Public health campaigns; Development and implementation of comprehensive women-specific confidential treatment models; and Empowering women. Specific goals in each of these areas that would achieve a positive impact on various aspects of the functioning of the current service delivery system for women with substance use disorders are suggested. Simultaneously seeking solutions in all four of these areas would improve the service delivery system and benefits women with substance use disorders.


Quebec Back Pain Disability Scale, Low Back Outcome Score and revised Oswestry low back pain disability scale for patients with low back pain due to degenerative disc disease: evaluation of Polish versions.
Evaluation and comparison of translated and culturally adapted self-reported measurements. The aim of this prospective study was to cross-culturally adapt the Polish versions of Revised Oswestry Disability Index (RODI-PL), Quebec Back Pain Disability Scale (QDS-PL), and the Low Back Outcome Score (LBOS-PL). The application of instruments in English, which have undergone translation must be subjected to validation studies. Such studies are necessary above all for instruments that have been adapted to establish their value and usefulness in studies of patient populations where English is not the native language. The translation was carried out according to International Quality of Life Association (IQOLA) Project and consisted of the following stages: translation, synthesis of the translations, back translation, expert committee, and testing of the prefinal versions of questionnaires. Eighty-five consecutive patients with low back pain due to spinal disc herniation and degenerative changes completed the QDS-PL, RODI-PL, LBOS-PL, and a Visual Analogue Scale twice within 2-day intervals. Mean duration of LBP was 45.9 months SD 55.5. The evaluation of degenerative changes in the lumbar region was carried out according to the Modic scale. Twenty-nine patients were categorized at type I, 4 patients were registered as type II, and 52 patients were type III. Cronbach α values for the LBOS-PL equaled 0.77, for the RODI-PL 0.85, and 0.95 for the QDS-PL. Item-total correlation confirmed that all scales are internally consistent. Test-retest reliability was excellent for RODI-PL and QDS-PL, but poor for LBOS-PL (0.88, 0.93, and 0.34, respectively). All questionnaires were significantly intercorrelated. We identified the strongest correlation between QDS-PL and RODI-PL (0.823, P < 0.001). The statistically significant correlation was identified between the QDS-PL and Modic Classification (rS = 0.226 P = 0.038). QBPDS-PL and RODI-PL are reliable and valid. Furthermore, investigation of the psychometric properties of the LBOS-PL in different spinal conditions is required. There is a relation between the results of the QDS-PL, and different levels of advancement in degenerative disease of the lumbar spine, according to the Modic Classification.


Cellular fatty acid composition of selected Pseudomonas species.
The cellular fatty acid composition of 10 reference strains representing eight species of Pseudomonas was determined by gas-liquid chromatography. A variety of acids were detected in these organisms, including branched and straight-chained acids, cyclopropane, and hydroxy acids. Comparison of the presence and relative amounts of these acids among strains was useful for distinguishing various Pseudomonas species.


[Indices of external respiration in patients with Hodgkin's disease long after irradiation of the mediastinum].
The function of the respiratory organs using the results of ventilation, gas exchange and roentgenopneumopolygraphy, was analysed in 98 Hodgkin's disease patients in a state of a prolonged remission (5-12 yrs.) after radiation therapy of the mediastinum. Pulmonary ventilation disorders were revealed in 43.9% of the patients. The degree of ventilation disorders depended on the degree of expression of radiation fibrosis in the paramediastinal regions of the lungs. Usually there were no clinical signs of the revealed changes.


Cholinesterase-Targeting microRNAs Identified in silico Affect Specific Biological Processes.
MicroRNAs (miRs) have emerged as important gene silencers affecting many target mRNAs. Here, we report the identification of 244 miRs that target the 3'-untranslated regions of different cholinesterase transcripts: 116 for butyrylcholinesterase (BChE), 47 for the synaptic acetylcholinesterase (AChE-S) splice variant, and 81 for the normally rare splice variant AChE-R. Of these, 11 and 6 miRs target both AChE-S and AChE-R, and AChE-R and BChE transcripts, respectively. BChE and AChE-S showed no overlapping miRs, attesting to their distinct modes of miR regulation. Generally, miRs can suppress a number of targets; thereby controlling an entire battery of functions. To evaluate the importance of the cholinesterase-targeted miRs in other specific biological processes we searched for their other experimentally validated target transcripts and analyzed the gene ontology enriched biological processes these transcripts are involved in. Interestingly, a number of the resulting categories are also related to cholinesterases. They include, for BChE, response to glucocorticoid stimulus, and for AChE, response to wounding and two child terms of neuron development: regulation of axonogenesis and regulation of dendrite morphogenesis. Importantly, all of the AChE-targeting miRs found to be related to these selected processes were directed against the normally rare AChE-R splice variant, with three of them, including the neurogenesis regulator miR-132, also directed against AChE-S. Our findings point at the AChE-R splice variant as particularly susceptible to miR regulation, highlight those biological functions of cholinesterases that are likely to be subject to miR post-transcriptional control, demonstrate the selectivity of miRs in regulating specific biological processes, and open new venues for targeted interference with these specific processes.


Activity, expression and function of a second Drosophila protein kinase A catalytic subunit gene.
The DC2 gene was isolated previously on the basis of sequence similarity to DC0, the major Drosophila protein kinase A (PKA) catalytic subunit gene. We show here that the 67-kD Drosophila DC2 protein behaves as a PKA catalytic subunit in vitro. DC2 is transcribed in mesodermal anlagen of early embryos. This expression depends on dorsal but on neither twist nor snail activity. DC2 transcriptional fusions mimic this embryonic expression and are also expressed in subsets of cells in the optic lamina, wing disc and leg discs of third instar larvae. A saturation screen of a small deficiency interval containing DC2 for recessive lethal mutations yielded no DC2 alleles. We therefore isolated new deficiencies to generate deficiency trans-heterozygotes that lacked DC2 activity. These animals were viable and fertile. The absence of DC2 did not affect the viability or phenotype of imaginal disc cells lacking DC0 activity or embryonic hatching of animals with reduced DC0 activity. Furthermore, transgenes expressing DC2 from a DC0 promoter did not efficiently rescue a variety of DC0 mutant phenotypes. These observations indicate that DC2 is not an essential gene and is unlikely to be functionally redundant with DC0, which has multiple unique functions during development.


B-cell antigen receptor expression and phosphatidylinositol 3-kinase signaling regulate genesis and maintenance of mouse chronic lymphocytic leukemia.
Chronic lymphocytic leukemia (CLL) is a frequent lymphoproliferative disorder of B cells. Although inhibitors targeting signal proteins involved in B-cell antigen receptor (BCR) signaling constitute an important part of the current therapeutic protocols for CLL patients, the exact role of BCR signaling, as compared to genetic aberration, in the development and progression of CLL is controversial. In order to investigate whether BCR expression per se is pivotal for the development and maintenance of CLL B cells, we used the TCL1 mouse model. By ablating the BCR in CLL cells from TCL1 transgenic mice, we show that CLL cells cannot survive without BCR signaling and are lost within 8 weeks in diseased mice. Furthermore, we tested whether mutations augmenting B-cell signaling influence the course of CLL development and its severity. The phosphatidylinositol-3-kinase (PI3K) signaling pathway is an integral part of the BCR signaling machinery and its activity is indispensable for B-cell survival. It is negatively regulated by the lipid phosphatase PTEN, whose loss mimics PI3K pathway activation. Herein, we show that PTEN has a key regulatory function in the development of CLL, as deletion of the Pten gene resulted in greatly accelerated onset of the disease. By contrast, deletion of the gene TP53, which encodes the tumor suppressor p53 and is highly mutated in CLL, did not accelerate disease development, confirming that development of CLL was specifically triggered by augmented PI3K activity through loss of PTEN and suggesting that CLL driver consequences most likely affect BCR signaling. Moreover, we could show that in human CLL patient samples, 64% and 81% of CLL patients with a mutated and unmutated IgH VH, respectively, show downregulated PTEN protein expression in CLL B cells if compared to healthy donor B cells. Importantly, we found that B cells derived from CLL patients had higher expression levels of the miRNA-21 and miRNA-29, which suppresses PTEN translation, compared to healthy donors. The high levels of miRNA-29 might be induced by increased PAX5 expression of the B-CLL cells. We hypothesize that downregulation of PTEN by increased expression levels of miR-21, PAX5 and miR-29 could be a novel mechanism of CLL tumorigenesis that is not established yet. Together, our study demonstrates the pivotal role for BCR signaling in CLL development and deepens our understanding of the molecular mechanisms underlying the genesis of CLL and for the development of new treatment strategies.


Forebrain-specific CRF overproduction during development is sufficient to induce enduring anxiety and startle abnormalities in adult mice.
Corticotropin releasing factor (CRF) regulates physiological and behavioral responses to stress. Trauma in early life or adulthood is associated with increased CRF in the cerebrospinal fluid and heightened anxiety. Genetic variance in CRF receptors is linked to altered risk for stress disorders. Thus, both heritable differences and environmentally induced changes in CRF neurotransmission across the lifespan may modulate anxiety traits. To test the hypothesis that CRF hypersignaling is sufficient to modify anxiety-related phenotypes (avoidance, startle, and conditioned fear), we induced transient forebrain-specific overexpression of CRF (CRFOE) in mice (1) during development to model early-life stress, (2) in adulthood to model adult-onset stress, or (3) across the entire postnatal lifespan to model heritable increases in CRF signaling. The consequences of these manipulations on CRF peptide levels and behavioral responses were examined in adulthood. We found that transient CRFOE during development decreased startle habituation and prepulse inhibition, and increased avoidance (particularly in females) recapitulating the behavioral effects of lifetime CRFOE despite lower CRF peptide levels at testing. In contrast, CRFOE limited to adulthood reduced contextual fear learning in females and increased startle reactivity in males but did not change avoidance or startle plasticity. These findings suggest that forebrain CRFOE limited to development is sufficient to induce enduring alterations in startle plasticity and anxiety, while forebrain CRFOE during adulthood results in a different phenotype profile. These findings suggest that startle circuits are particularly sensitive to forebrain CRFOE, and that the impact of CRFOE may be dependent on the time of exposure.